Antioxidative and Antimycotoxigenic Efficacies of Thunbergia laurifolia Lindl. for Addressing Aflatoxicosis in Cherry Valley Ducks.

This study aimed to assess the effectiveness of aflatoxin B1 (AFB1) and Thunbergia laurifolia extract (TLE) in the diets of Cherry Valley ducklings. Our investigation covered growth indicators, blood biochemical indices, meat quality, intestinal morphology, immune response, and CP450 enzyme-related gene expression. We conducted the study with 180 seven-day-old Cherry Valley ducks, randomly divided into five dietary treatments. These treatments included a basal diet without AFB1 (T1 group), TLE, or a commercial binder; the basal diet containing 0.1 mg AFB1/kg (T2 group), 0.1 mg AFB1/kg and 100 mg TLE/kg (T3 group), 0.1 mg AFB1/kg and 200 mg TLE/kg (T4 group), and 0.1 mg AFB1/kg and 0.5 g/kg of a commercial binder (T5 group), respectively. Ducklings fed with the T2 diet exhibited lower final body weight (BW), average body weight gain (ADG), and poor feed conversion ratio (FCR) during the 42-day trials. However, all ducklings in the T3, T4, and T5 groups showed significant improvements in final BW, ADG, and FCR compared to the T2 group. Increased alanine transaminase (ALT) concentration and increased expression of CYP1A1 and CYP1A2 indicated hepatotoxicity in ducklings fed the T2 diet. In contrast, ducklings fed T3, T4, and T5 diets all showed a decrease in the expression of CYP1A1 and CYP1A2, but only the T4 treatment group showed improvement in ALT concentration. AFB1 toxicity considerably raised the crypt depth (CD) in both the duodenum and jejunum of the T2 group, while the administration of 200 mg TLE/kg (T4) or a commercial binder (T5) effectively reduced this toxicity. Additionally, the villus width of the jejunum in the T2 treatment group decreased significantly, while all T3, T4, and T5 groups showed improvement in this regard. In summary, T. laurifolia extract can detoxify aflatoxicosis, leading to growth reduction and hepatic toxicosis in Cherry Valley ducklings.

Tenuazonic acid-induced mycotoxicosis in an immunosuppressed mouse model and its prophylaxis with cinnamaldehyde.

Patients with impaired immune systems are particularly vulnerable to infections. With the increasing number of immunocompromised patients, it becomes necessary to design studies that evaluate the effects of toxic contaminants that are a part of our daily lives. Simultaneously, the management of these toxic components also becomes essential. Therefore, the present study evaluated the possible protective role of cinnamaldehyde (Cin) against tenuazonic acid-induced mycotoxicosis in the immunosuppressed murine model. Tenuazonic acid (TeA), a toxin usually produced by Alternaria species, is a common contaminant in tomato and tomato-based products. Evaluating the potential toxicity of a hazardous chemical necessitates the use of in vitro, in vivo, and in silico methods. Here, the immunomodulatory effect of TeA was assessed in vitro using mouse splenocytes. In silico docking was carried out for the tumour markers of eight organs and TeA. The haematological, histopathological, and biochemical aspects were analysed in vivo. The sub-chronic intoxication of mice with TeA showed elevated malondialdehyde, reduced catalase, and superoxide dismutase production, along with abnormal levels of aspartate aminotransferase and alanine transaminase. The treatment with Cin prevented TeA-induced alterations of antioxidant defense enzyme activities and significantly forbade TeA-induced organ damage, showing therapeutic effects and toxicity reduction in TeA-induced mycotoxicosis.

Nanosilver effects on growth parameters in experimental aflatoxicosis in broiler chickens.

Aflatoxicosis is a cause of economic losses in broiler production. In this study, the effect of one commercial nanocompound, Nanocid (Nano Nasb Pars Co., Iran) was evaluated in reduction of aflatoxin effects on the growth and performance indices in broiler chickens suffering from experimental aflatoxicosis. For this, a total of 300 one-day-old broiler chicks (Ross strain) were randomly divided into 4 groups with 3 replicates of 15 chicks in each separated pen during the 28-day experiment. Treatment groups including group A: chickens fed basal diet, group B: chickens fed 3 ppm productive aflatoxin in basal diet, group C: chickens fed basal diet plus 2500 ppm Nanocid, and group D: chickens fed 3 ppm productive aflatoxin and 2500 ppm Nanocid, in basal diet. Data on body weight, body weight gain (BWG), feed intake, and feed conversion ratio (FCR) were recorded at weekly intervals. Also cumulative data were assessed. Results showed, although supplement of Nanocid to conventional diet had no effect on performance but addition of Nanocid to diet containing 3 ppm aflatoxin increased significantly the cumulative BWG, cumulative feed consumption and decreased FCR in the last 2 weeks of experimental period. The improvement in these performance indices by supplement of Nanocid to diet containing aflatoxin showed the ability of Nanocid to diminish the inhibitory effects of aflatoxin.

Investigation of a Novel Multicomponent Mycotoxin Detoxifying Agent in Amelioration of Mycotoxicosis Induced by Aflatoxin-B1 and Ochratoxin A in Broiler Chicks.

The present study was designed to determine the efficacy of a novel multicomponent mycotoxin detoxifying agent (MMDA) containing modified zeolite (Clinoptilolite), Bacillus subtilis, B. licheniformis, Saccharomyces cerevisiae cell walls and silymarin against the deleterious effects of Aflatoxin B1 (AFB1) and Ochratoxin A (OTA) in broiler chicks. A total of 160 one-day-old Ross 308® broiler chicks were randomly allocated in four treatment groups, with four replicates, according to the following experimental design for 42 days. Group A received a basal diet; Group B received a basal diet contaminated with AFB1 and OTA at 0.1 mg/kg and 1 mg/kg, respectively; Group C received a basal diet contaminated with AFB1 and OTA and MMDA at 1 g/kg feed, and Group D received a basal diet contaminated with AFB1 and OTA and MMDA at 3 g/kg feed. Results showed that ingested mycotoxins led to significant (p ≤ 0.05) reduction in body weight and feed conversion from 25 days of age, induced histopathological changes, increased the pH of the intestinal content, and altered the biochemical profile of birds with significantly (p ≤ 0.05) increased aspartate aminotransferase (AST) values (p ≤ 0.05). On the other hand, the supplementation of MMDA significantly (p ≤ 0.05) improved the feed conversion ratio (FCR) during the second part of the study, diminished biochemical alterations, reduced pH in jejunal and ileal content, and E. coli counts in the caeca of birds (p ≤ 0.05). It may be concluded that the dietary supplementation of the MMDA partially ameliorated the adverse effects of AFB1 and OTA in broilers and could be an efficient tool in a mycotoxin control program.

Molecular mechanisms of fumonisin B1-induced toxicities and its applications in the mechanism-based interventions.

Fumonisin B1 (FB1), the most toxic member of fumonisins, is one of the most common mycotoxins contaminating feed and food. It has been shown to produce pleiotropic toxicities in animals including neurotoxicity, hepatotoxicity and nephrotoxicity, and with highly potential impact on human health. Proposed mechanisms include disruption of sphingolipid metabolism, induction of oxidative stress, activation of endoplasmic reticulum (ER) stress and MAPKs, modulation of autophagy and alteration of DNA methylation. These mechanistic findings provide the theoretical basis for effectively managing FB1-induced adverse health effects. In this review, we will summarize the present understanding of the molecular basis underlying FB1-mediated toxicities, discuss the feasibility of developing mechanism-based approach to prevent FB1-induced toxicities, and suggest the issues that need to be addressed in the future in the field of FB1 toxicity-related studies.

The effects of astaxanthin on liver histopathology and expression of superoxide dismutase in rat aflatoxicosis.

The metabolism of aflatoxin B1 (AFB1) generates reactive oxygen species (ROS) that destroys hepatocytes. Meanwhile, astaxanthin (AX) is known to have stronger antioxidative activity than other carotenoids. This study aimed to investigate hepatoprotective role of AX from AFB1-induced toxicity in rat by histopathological study and immunohistochemistry of Cu/Zn-SOD (SOD1) which acts as the first enzyme in antioxidative reaction against cell injury from ROS. Twenty Wistar rats were randomly divided into 4 groups. The control and AFB1 groups were gavaged by water for 7 days followed by a single DMSO and 1 mg/kg AFB1, respectively. The AXL+ AFB1 and AXH+ AFB1 groups were given of 5 mg/kg and 100 mg/kg AX for 7 days before 1 mg/kg AFB1 administration. The result showed significantly elevated liver weight per 100 g body weight in AFB1 group. The histopathological finding revealed vacuolar degeneration, necrosis, megalocytosis and binucleation of hepatocytes with bile duct hyperplasia in AFB1 group. The severities of pathological changes were sequentially reduced in AXL+AFB1 and AXH+AFB1 groups. Most rats in AXH+AFB1 group owned hypertrophic hepatocytes and atypical proliferation of cholangiocytes which are adaptive responses to severe hepatocyte damage. The SOD1 expression was also significantly higher in AXH+AFB1 group than solely treated AFB1 and AXL+AFB1 groups. In conclusion, AX alleviated AFB1-induced liver damage in rat by stimulating SOD1 expression and transdifferentiation of cholangiocytes in dose dependent manner.

Clinico-pathomorphological, serum biochemical and histological studies in broilers fed ochratoxin A and a toxin deactivator (Mycofix Plus).

1. Toxic effects of two concentrations (0.5 and 1 mg/kg) of ochratoxin A (OTA) and attenuating effects of a toxin deactivator (Mycofix Plus(MTV INSIDE)) containing the yeast Trichosporon mycotoxinivorans on the performance (feed conversion ratio; body weight gain), serum enzymes (lactate dehydrogenase, gamma-glutamyltranspeptidase and aspartate aminotransferase) and clinico-pathomorphology of internal organs were studied in 270 one-day-old broiler chicks divided into 9 groups over a 42-d period. 2. Feed conversion ratios (FCR) in groups fed toxin deactivator were improved compared with groups receiving OTA only. An increase in the relative weight of kidney and liver was observed in groups fed 0.5 and 1 mg/kg OTA on day 42 of the experiment as compared with the control group. In contrast, relative weights of bursa of Fabricius and spleen were not significantly affected in experimental groups exposed to OTA as compared to control groups determined on days 28 and 42 of age. 3. Serum enzymes (LDH, GGT and AST) values in OTA treated groups determined on days 28 and 42 were higher than those of the control group. 4. Histopathological examination of kidney on day 42 revealed degenerative changes in the epithelial cells of the proximal convoluted tubules and massive necrosis of the proximal tubular epithelial cells. These changes were less marked in birds receiving 0.5 mg/kg OTA than in those receiving 1 mg/kg. In general, histological changes in kidneys, liver, bursa and spleen were less pronounced in birds receiving OTA and toxin deactivator concomitantly. 5. Dietary OTA at 0.5 and 1 mg/kg adversely affects FCR, increases the serum liver enzymes and induces pronounced pathomorphological and histological changes in internal organs of broiler chicks. Co-administration of OTA with deactivator attenuated the harmful effects.

[Effect of probiotic preparation based on Bacillus subtilis (BPS-44) in experimental mycotoxicoses of chickens].

When Road-Island breed chickens were given fodder which included toxin in concentration of 16 mg/kg or T-2 toxin in concentration of 10 Mg/kg, that resulted in the decrease of the live weight, increase in the relative weight of the liver, kidneys, pancreas and heart, as well as the decrease of concentration of Bacillus genus bacteria in the caecum and rectum content compared with the control group chickens. No distinctions were observed in activity of alanine aminotransferase and concentration of total protein in the blood plasm. The drinking of probiotic preparation BPS-44 when feeding with forage contaminated by HT-2 or T-2 toxin resulted in the increase of the live weight, normalization of relative weights of viscera, increase in concentration of Bacillus genus bacteria in the intestine compared with chickens which received only mycotoxins.

Acute aflatoxicosis: case report.

The objective of this presentation is to document the salient clinical findings in a case of aflatoxicosis and to review the literature on the same so as to increase the index of suspicion, enhance early diagnosis and improve management. The case was a 17-year-old schoolboy presenting with vomiting, features of infection and gastrointestinal tract symptoms. Examination revealed a very ill looking pale patient with abdominal distension, tenderness and rectal bleeding and easy bruisability. Investigations showed abnormal liver function tests, pancytopenia and elevated serum levels of aflatoxins. Management consisted of supportive care including antibiotics and antifungal therapy, transfusion of red blood cells and fresh frozen plasma. His recovery was uneventful. The literature on human aflatoxicosis shows that the presentation may be acute, subacute and chronic. The degree of emanating clinical events also conforms to status of the aflatoxicosis. Overall, the features are protean and may masquerade many other forms of toxaemias. In conclusion, the diagnosis of aflatoxicosis takes cognisance of geographical location, past events, staple diet and clinical features to exclude other infections. Also required are high index of suspicion and importantly serum levels of aflatoxin. Treatment strategies involved use of antimicrobials and supporting the damaged multi-organs.

Protective effect of L-carnitine against oxidative damage caused by experimental chronic aflatoxicosis in quail (Coturnix coturnix).

This study was designed to evaluate the effect of L-carnitine supplementation on the plasma malondialdehyde (MDA) and whole blood reduced glutathione (GSH) concentrations in experimentally-induced chronic aflatoxicosis in quails. For this purpose, a total of 80 quails up to 8 weeks old were divided into four equal groups. Group I served as control, Group II was given L-carnitine at the dose of 200 mg/litre in the drinking water for 60 days, Group III was given 60 microg total aflatoxin/kg diet for 60 days, and Group IV was given both 60 microg total aflatoxin/kg diet and 200 mg L-carnitine/litre in the drinking water for 60 days. Aflatoxin treatment caused a significant increase in plasma MDA and a significant decrease in blood GSH concentrations. On the other hand, there was a significant decrease in plasma MDA and a significant increase in whole blood GSH in the L-carnitine-supplemented group. The present study demonstrated that L-carnitine brought about the inhibition of lipid peroxidation by enhancing antioxidant capacity in quails with chronic aflatoxicosis.

Hydrated sodium calcium aluminosilicate (HSCAS), acidic HSCAS, and activated charcoal reduce urinary excretion of aflatoxin M1 in turkey poults. Lack of effect by activated charcoal on aflatoxicosis.

In one experiment, the effect of inorganic sorbents on the metabolic fate of aflatoxin B1 (AFB1) was studied in turkey poults. At 5 weeks of age, female poults were surgically colostomized and 9 days later orally dosed with 0.75 mg AFB1/kg BW. Hydrated sodium calcium aluminosilicate (HSCAS), acidic HSCAS, and activated charcoal (AC) were tested, by concomitant administration with AFB1. Urine was collected up to 48 h post-dosing and analyzed for aflatoxin M1 (AFM1) which was the major metabolite found in all treatment groups. Hydrated sodium calcium aluminosilicate, previously proven beneficial in alleviating aflatoxicosis in farm animals, reduced urinary AFM1 output when orally dosed simultaneous with AFB1. Also, acidic HSCAS and AC significantly decreased AFM1 excretion when administered concomitantly with AFB1. A second experiment was conducted to evaluate the ability of two types of AC to modify aflatoxicosis when added to aflatoxin (AF)-contaminated (from culture material) diets of turkey poults. Although AC was able to decrease AFM1 excretion in the first experiment, no protective effects from AF toxicity were observed in the feeding study.

Ameliorative effects of dietary clinoptilolite on pathological changes in broiler chickens during aflatoxicosis.

The amelioration of aflatoxicosis in broiler chickens was examined by feeding two concentrations of natural zeolite (clinoptilolite). Clinoptilolite (ClI), incorporated into the diet at 1.5 and 2.5 per cent, was evaluated for the ability to reduce the deleterious effects of 2.5 mg total aflatoxin (AF) kg(-1)diet on growing broiler chicks from 1 to 21 days of age. A total of 360 broiler chicks were divided into six treatment groups [Control, AF, CLI (1.5 per cent), AF plus CLI (1.5 per cent), CLI (2.5 per cent), and AF plus CLI (2.5 per cent)] each consisting of 60 chicks. Compared to controls, the AF consuming chicks showed increases in the relative weights of liver and kidney; and gross-histopathologic hepatic lesions such as paleness, friability, diffuse hydropic degeneration and/or fatty change, bile-duct hyperplasia and periportal fibrosis. Glumerular hypertrophy, increases in the number of mesengial cells and hydropic degeneration of tubuler epithelium in kidneys of chicks fed diet AF alone were also observed. Atrophy and lymphoid depletion were seen in the thymuses and bursa of Fabricius from the chicks fed AF alone. The additions of CLI (1.5 and 2.5 per cent) to the AF -containing diet moderately (significantly in some cases) decreased the number of affected broilers and/or the severity of lesions. The addition of CLI to the AF-free diet did not produce any significant changes compared with the controls. These results suggest that CLI was effective for the protection of AF-toxication in broilers and it could contribute to a solution of the AF problem in poultry production.

Influence of a hydrated sodium calcium aluminosilicate and virginiamycin on aflatoxicosis in broiler chicks.

Male broiler chicks were fed diets containing 0 or 3.5 mg aflatoxin (AF)/kg from 1 to 28 d of age. The diets were amended with either .5% hydrated sodium calcium aluminosilicate (HSCAS), 16.5 mg virginiamycin (VM)/kg of diet, or .5% HSCAS + 16.5 mg VM/kg of diet to determine the effects of these compounds during aflatoxicosis. When compared with controls (814 g), BW gains were significantly lower for broilers fed the AF (731 g) or AF+VM (716 g) diets. No differences were found between the BW gains of chicks fed diets without aflatoxin (814 g) and those of chicks fed AF+HSCAS (793 g) or AF+HSCAS+VM (803 g), indicating approximately 75% protection against decreased BW gain by HSCAS and 87% protection by the HSCAS+VM combination. Relative weights of the liver and kidney and creatine kinase activity were significantly increased and albumin, total protein, cholesterol, uric acid, and inorganic phosphorus concentrations were decreased in chicks fed the diet containing AF alone. With the exception of albumin, HSCAS was effective in restoring these values to control values. The findings of this research suggest that HSCAS and HSCAS+VM can counteract some of the toxic effects of AF in growing broiler chicks.

Effect of tryptophan supplementation on aflatoxicosis in laying hens.

Two experiments were conducted to determine the relationship of supplemental Trp on liver fat accumulation and egg production during aflatoxicosis in laying hens. In Experiment 1, two levels of Trp (0 and 2,000 ppm; basal = .16% Trp) and two levels of aflatoxin (AFLA) (0 and 10 ppm) were supplemented to a complete layer ration. In Experiment 2, a third level of AFLA (5 ppm) was added to the design. Single Comb White Leghorn hens (58 and 68 wk old) were fed the diets for 3 wk for Experiments 1 and 2, respectively. Henday production and egg weights were measured daily. Feed intake was measured weekly. Liver weights, liver moisture, and liver total lipids were determined at the end of each trial. In Experiment 1, supplemental Trp by itself caused a significant (P less than .01) reduction in total liver lipids compared to the controls (no Trp or AFLA). Adding trp and AFLA increased total liver lipids and caused a significant (P less than .05) decrease in egg production compared with adding AFLA alone. Total liver lipids were 41.1, 32.8, 54.8, and 62.4% (dry weight basis) for 0 Trp:0 AFLA, 2,000 Trp:0 AFLA, 0 Trp:10 AFLA, and 2,000 Trp:10 AFLA, respectively. Similar results were observed in Experiment 2. It was concluded that supplemental Trp by itself caused a reduction in total liver lipids, but when supplemented to a diet containing AFLA, Trp caused an increased severity of lesions associated with aflatoxicosis in layers.

Lipoic acid as a potential first agent for protection from mycotoxins and treatment of mycotoxicosis.

Mycotoxins--toxic substances produced by fungi or molds--are ubiquitous in the environment and are capable of damaging multiple biochemical mechanisms, resulting in a variety of human symptoms referred to collectively as "mycotoxicosis." In fact, mycotoxins mimic multiple xenobiotics, not only with respect to their ultimate damage, but also in their routes of detoxification. This suggests potential therapeutic options for the challenging treatment of mycotoxicosis. In this brief review, the author examines the use of lipoic acid as an example of an inexpensive and available nutrient that has been shown to protect against, or reverse, the adverse health effects of mycotoxins.

Experimental mycotoxicosis in chickens induced by ochratoxin A and penicillic acid and intervention with natural plant extracts.

The combined toxic effect of ochratoxin A (OTA) and penicillic acid (PA) on the body mass, the weight and pathomorphology of some internal organs was studied in 85 broiler chickens fed a mouldy diet containing 130, 300 or 800 ppb OTA and 1000-2000 ppb PA. The main pathomorphological changes were cloudy swelling and granular degeneration in the epithelium and mononuclear cell proliferation and activation of capillary endothelium in the kidney and liver; degenerative changes and depletion of lymphoid cells in lymphoid organs (bursa of Fabricius, thymus and spleen) were also seen. Protective effects of 5% total water extract of artichoke and a new natural phytosubstance Rosallsat against these pathomorphological changes were observed. A significant decrease in body mass and relative weight of lymphoid organs was found after 6 weeks of exposure and a greater decrease after 10 weeks of exposure to OTA and PA, and a protective effect of artichoke extract and a slight effect of Rosallsat against that decrease was observed. A significant increase in relative weight of liver and kidneys was also observed as well as a protective effect of artichoke extract against that increase. The quantity of OTA and the percentage of positive samples were significantly lower in tissues of chickens treated with artichoke extract or Rosallsat in addition to OTA than in those treated with only OTA.

Dietary glutamate supplementation ameliorates mycotoxin-induced abnormalities in the intestinal structure and expression of amino acid transporters in young pigs.

The purpose of this study was to investigate the hypothesis that dietary supplementation with glutamic acid has beneficial effects on growth performance, antioxidant system, intestinal morphology, serum amino acid profile and the gene expression of intestinal amino acid transporters in growing swine fed mold-contaminated feed. Fifteen pigs (Landrace×Large White) with a mean body weight (BW) of 55 kg were randomly divided into control group (basal feed), mycotoxin group (contaminated feed) and glutamate group (2% glutamate+contaminated feed). Compared with control group, mold-contaminated feed decreased average daily gain (ADG) and increased feed conversion rate (FCR). Meanwhile, fed mold-contaminated feed impaired anti-oxidative system and intestinal morphology, as well as modified the serum amino acid profile in growing pigs. However, supplementation with glutamate exhibited potential positive effects on growth performance of pigs fed mold-contaminated feed, ameliorated the imbalance antioxidant system and abnormalities of intestinal structure caused by mycotoxins. In addition, dietary glutamate supplementation to some extent restored changed serum amino acid profile caused by mold-contaminated feed. In conclusion, glutamic acid may be act as a nutritional regulating factor to ameliorate the adverse effects induced by mycotoxins.

Intoxicación por Chlorophyllum molybdites: Presentación de dos casos y revisión de la literatura / Intoxication by Chlorophyllum molybdites: Presentation of two cases and review of the literature

La intoxicación por hongos, especialmente con fines alimenticios, es un importante problema terapéutico. El hongo Chlorophyllum molybdites, es uno de los principales agentes causantes de intoxicación en países Latinoameri-canos, debido a su similitud con el hongo Agaricus comestible. Se presentan dos casos de intoxicación por Chlorophyllum molybdites los cuales fueron ingeridos y posteriormente desarrollaron manifes-taciones clínicas. Se presenta con su revisión bibliográfica centrada en las diferentes opciones terapéuticas...(AU)

Aquilegia vulgaris L. extract counteracts oxidative stress and cytotoxicity of fumonisin in rats.

Exposure to fumonisins (FB) is known to have toxic and carcinogenic effects in different animal species, and to express toxicity in cells via the induction of oxidative stress. The aim of the current study was to evaluate the protective effects of the ethanol extract of Aquilegia vulgaris L. against the oxidative stress and the genotoxicity using micronucleus assay and random amplified polymorphism DNA (RAPD-PCR) in FB-treated rats. Sixty mature female Sprague-Dawley were divided into six treatment groups and treated for 4 weeks as follow: the control group, the group fed FB-contaminated diet (200 mg/kg diet), the groups treated orally with the extract (5 and 10 mg/kg bw) and the groups fed FB-contaminated diet and treated with the extract at the two doses. The results showed that treatment with FB alone disturbed lipid profile in serum, increases Sa/So ratio, induces micronucleated polychromatic erythrocytes (Mn-PCEs) in bone marrow, increases DNA and RNA in liver accompanied with significant changes in histological picture The extract alone at the two tested doses did not induce any significant changes in the biochemical or histological picture. The combined treatment showed significant improvements in all biochemical, cytogenetic parameters tested and histological pictures in the liver tissues. Moreover, this improvement was more pronounced in the group received the high dose of the extract. It could be concluded that the ethanol extract of A. vulgaris induced its protective effect via the increase in the antioxidant capacity, inhibition of lipid peroxidation and scavenging of free radicals.